Effects of Increased NADPH Concentration by Metabolic Engineering of the Pentose Phosphate Pathway on Antibiotic Production and  Sporulation in Streptomyces lividans TK24

Jin, Xue-Mei; Chang, YongKeun; Lee, Jae Hag; Hong, Soon-Kwang

doi:10.4014/jmb.1707.07046

Cited by 16 publications

(5 citation statements)

References 22 publications

(41 reference statements)

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“…Consequently, a reduced generation of NADPH in SC is predicted to lead to high oxidative stress that was proposed to be an important trigger of ACT biosynthesis in SC ( Esnault et al, 2017 ; Virolle, 2020 ). Consistently, reports in the literature mentioned that a reduced carbon flux through PPP, a consequence of the deletion of genes encoding the first enzymes of the PPP, the isoenzymes glucose 6-P-dehydrogenase (zwf1/SCO6661 zwf2/SCO1937), and/or the 6-phosphogluconolactonase (devD/SCO1939), resulted in increased ACT levels ( Butler et al, 2002 ), whereas conversely, the over-expression of these enzymes led to reduced ACT levels ( Jin et al, 2017 ). At last, it should be stressed that, the abundance pattern of enzymes of the PPP being similar in the two SL strains, a lower activity of the PPP cannot account for a higher oxidative stress triggering ACT production in the pptA mutant.…”

Section: Resultssupporting

confidence: 66%

A Proteomic Analysis Indicates That Oxidative Stress Is the Common Feature Triggering Antibiotic Production in Streptomyces coelicolor and in the pptA Mutant of Streptomyces lividans

et al. 2022

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In most Streptomyces species, antibiotic production is triggered in phosphate limitation and repressed in phosphate proficiency. However, the model strain, Streptomyces coelicolor, escapes this general rule and produces actinorhoddin (ACT), a polyketide antibiotic, even more abundantly in phosphate proficiency than in phosphate limitation. ACT was shown to bear “anti-oxidant” properties suggesting that its biosynthesis is triggered by oxidative stress. Interestingly, Streptomyces lividans, a strain closely related to S. coelicolor, does not produce ACT in any phosphate condition whereas its pptA/sco4144 mutant produces ACT but only in phosphate limitation. In order to define the potentially common features of the ACT producing strains, these three strains were grown in condition of low and high phosphate availability, and a comparative quantitative analysis of their proteomes was carried out. The abundance of proteins of numerous pathways differed greatly between S. coelicolor and the S. lividans strains, especially those of central carbon metabolism and respiration. S. coelicolor is characterized by the high abundance of the complex I of the respiratory chain thought to generate reactive oxygen/nitrogen species and by a weak glycolytic activity causing a low carbon flux through the Pentose Phosphate Pathway resulting into the low generation of NADPH, a co-factor of thioredoxin reductases necessary to combat oxidative stress. Oxidative stress is thus predicted to be high in S. coelicolor. In contrast, the S. lividans strains had rather similar proteins abundance for most pathways except for the transhydrogenases SCO7622-23, involved in the conversion of NADPH into NADH. The poor abundance of these enzymes in the pptA mutant suggested a deficit in NADPH. Indeed, PptA is an accessory protein forcing polyphosphate into a conformation allowing their efficient use by various enzymes taking polyphosphate as a donor of phosphate and energy, including the ATP/Polyphosphate-dependent NAD kinase SCO1781. In phosphate limitation, this enzyme would mainly use polyphosphate to phosphorylate NAD into NADP, but this phosphorylation would be inefficient in the pptA mutant resulting in low NADP(H) levels and thus high oxidative stress. Altogether, our results indicated that high oxidative stress is the common feature triggering ACT biosynthesis in S. coelicolor and in the pptA mutant of S. lividans.

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Section: Resultssupporting

confidence: 66%

A Proteomic Analysis Indicates That Oxidative Stress Is the Common Feature Triggering Antibiotic Production in Streptomyces coelicolor and in the pptA Mutant of Streptomyces lividans

et al. 2022

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“…Most of the biosynthetic pathways for secondary metabolites are influenced by carbon metabolism and supply of cytosolic NADPH [ 40 ]. NADPH is mainly synthesized by a central carbon metabolic pathway, which can be strengthened by redistributing the glycolysis flux to the pentose phosphate pathway [ 41 ].…”

Section: Resultsmentioning

confidence: 99%

Engineering of Biosynthesis Pathway and NADPH Supply for Improved L-5-Methyltetrahydrofolate Production by Lactococcus lactis

Liu

et al. 2021

J. Microbiol. Biotechnol.

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L-5-methyltetrahydrofolate (5-MTHF) is one of the biological active forms of folate, which is widely used as a nutraceutical. However, low yield and serious pollution associated with the chemical synthesis of 5-MTHF hampers its sustainable supply. In this study, 5-MTHF production was improved by engineering the 5-MTHF biosynthesis pathway and NADPH supply in Lactococcus lactis for developing a green and sustainable biosynthesis approach. Specifically, overexpressing the key rate-limiting enzyme methylenetetrahydrofolate reductase led to intracellular 5-MTHF accumulation, reaching 18 μg/l. Next, 5-MTHF synthesis was further enhanced by combinatorial overexpression of 5-MTHF synthesis pathway enzymes with methylenetetrahydrofolate reductase, resulting in 1.7-fold enhancement. The folate supply pathway was strengthened by expressing folE encoding GTP cyclohydrolase I, which increased 5-MTHF production 2.4-fold to 72 μg/l. Furthermore, glucose-6-phosphate dehydrogenase was overexpressed to improve the redox cofactor NADPH supply for 5-MTHF biosynthesis, which led to a 60% increase in intracellular NADPH and a 35% increase in 5-MTHF production (97 μg/l). To reduce formation of the by-product 5-formyltetrahydrofolate, overexpression of 5-formyltetrahydrofolate cyclo-ligase converted 5-formyltetrahydrofolate to 5,10-methyltetrahydrofolate, which enhanced the 5-MTHF titer to 132 μg/l. Finally, combinatorial addition of folate precursors to the fermentation medium boosted 5-MTHF production, reaching 300 μg/l. To the best of our knowledge, this titer is the highest achieved by L. lactis . This study lays the foundation for further engineering of L. lactis for efficient 5-MTHF biosynthesis.

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“…Furthermore, one allosteric effector gene, opc, which is required for ZWF activity, was found to be overexpressed in various combinations. The final recombinant strain, S. lividans TK24/pWHM3-Z23O2, showed improved NADPH production and suggested that metabolic engineering of PPP routes is one of the suitable pathways for secondary metabolite production like antibiotics (Jin et al, 2017).…”

Section: Metabolic Glycoengineering Emerged As a Novel Antibacterial ...mentioning

confidence: 99%

Bacterial glycobiotechnology: A biosynthetic route for the production of biopharmaceutical glycans

Paliya

Sharma

Tuohy

et al. 2023

Biotechnology Advances

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Effects of Increased NADPH Concentration by Metabolic Engineering of the Pentose Phosphate Pathway on Antibiotic Production and Sporulation in Streptomyces lividans TK24

Cited by 16 publications

References 22 publications

A Proteomic Analysis Indicates That Oxidative Stress Is the Common Feature Triggering Antibiotic Production in Streptomyces coelicolor and in the pptA Mutant of Streptomyces lividans

A Proteomic Analysis Indicates That Oxidative Stress Is the Common Feature Triggering Antibiotic Production in Streptomyces coelicolor and in the pptA Mutant of Streptomyces lividans

Engineering of Biosynthesis Pathway and NADPH Supply for Improved L-5-Methyltetrahydrofolate Production by Lactococcus lactis

Bacterial glycobiotechnology: A biosynthetic route for the production of biopharmaceutical glycans

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